Iterative multitarget evolution dramatically enhances the enantioselectivity and catalytic efficiency of Bacillus subtilis esterase towards bulky benzoate esters ofdl-menthol

Abstract

Structure-based directed evolution has been successfully applied to Bacillus subtilis esterase to produce a mutant with higher enantioselectivity and elevated efficiency.

Multiparameter Optimization in Directed Evolution: Engineering Thermostability, Enantioselectivity, and Activity of an Epoxide Hydrolase

“…However, these additional properties are generally not improved in the process of thermostabilization. Various approaches to the optimization of two parameters in specific cases have been reported, but general guidelines remain to be issued. − , In most cases substantial experimental efforts had to be invested. Two general options are possible: sequential or simultaneous optimization of two or more parameters.…”

Production of β‐Alanine from Fumaric Acid Using a Dual‐Enzyme Cascade

“…These two methods complement one another and have been used together to overcome the limitations of natural enzymes in enzyme cascades ,. For example, Gong and colleagues dramatically enhanced catalytic efficiency of esterase by combining multi‐target evolution (including site‐directed mutagenesis and random mutagenesis) as a way to enhance L‐menthol synthesis from DL‐menthyl benzoate; (ii) The second group, regulating enzyme expression, can be accomplished by optimization at the transcriptional or translational levels. Regulation can be accomplished through a number approaches such as promoter replacement, RBS regulation, multi‐plasmid systems, and gene copies modification.…”

Multidisciplinary approaches for enzyme biocatalysis in pharmaceuticals: protein engineering, computational biology, and nanoarchitectonics

“…Further exemplification of enzyme engineering can be found in Table 2, where additional instances are provided. [122][123][124][125][126][127][128][129][130][131][132][133][134][135][136][137][138][139][140] Enhanced catalytic activity accelerates reactions, stability ensures that enzymes can endure challenging environments, and refined substrate specificity opens doors to a plethora of novel applications. However, these methods still have several limitations, for example, directed evolution can be timeconsuming and resource-intensive and may not always yield enzymes with the desired properties.…”